Methods and compositions for enhancing or maintaining fertility

ABSTRACT

Embodiments of the invention generally relate to methods and supplements for increasing, enhancing, or maintaining fertility in a human being.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of application Ser. No.14/301,219, filed Jun. 10, 2014, which is a continuation/divisionalapplication of U.S. Pat. 8,747,922, issued on Jun. 10, 2014, which willissue as U.S. Patent the disclosure of which is hereby incorporatedherein in its entirety by this reference.

FIELD OF THE DISCLOSURE

The present disclosure generally relates to methods and supplements forimproving fertility in a subject. In some embodiments, a nutritionalsupplement may be administered to a subject to increase levels of growthhormone (hGH) in the subject, thereby increasing fertility in thesubject.

BACKGROUND

The primary biological function of human growth hormone (hGH) includesstimulating growth, cell repair, and regeneration. Once the primarygrowth period of adolescence concludes, the primary function of hGH inadulthood becomes that of cell regeneration and repair, helpingregenerate skin, bones, heart, lungs, liver, and kidneys to theiroptimal, youthful cell levels. As is the case with many of our otherhormones or their precursors, such as testosterone, estrogen,progesterone, DHEA, and melatonin, hGH levels decline with age.Therapeutically, many of these hormones can be replaced to offset someof the effects of aging such as menopausal symptoms in women or erectiledysfunction in men. Additionally, hGH and testosterone affect sterilityand fertility in animals, including human beings.

The human body, like every other living entity, works on daily, orcircadian, as well as monthly and annual rhythms. Daily growth hormonesecretion diminishes with age with roughly half the levels at age 40that we had when we were 20, and about one-third of those youthfullevels at age 60. In some 60-year-olds, the levels are as low as 25% ofthe hGH levels in a 20-year-old. Symptoms of aging include loss ofmuscle, increase of fat, decreased physical mobility, decreased energylevels, and as a result, diminished socialization, diminished healingability, and an increased risk of cardiovascular disease and decreasedlife expectancy. Low hGH levels are associated with the aging processand early onset of disease. For example, Rosen and Bengtsson noted anincreased death rate from cardiovascular disease in hGH deficientpatients. (Rosen, T., Bengtsson, B. A., Lancet 336 (1990): 285-2880)

Low hGH has been shown to decrease fertility in humans. See, e.g.,Homburg, R., A. Singh, et al. (2012), “The re-growth of growth hormonein fertility treatment: a critical review.” Hum Fertil (Camb) 15(4):190-193; Giampietro, A., D. Milardi, et al. (2009), “The effect oftreatment with growth hormone on fertility outcome in eugonadal womenwith growth hormone deficiency: report of four cases and review of theliterature.” Fertil Steril 91(3): 930 e937-911; Kalra, S., B. Kalra, etal. (2008), Growth hormone improves semen volume, sperm count andmotility in men with idopathic normogonadotropic infertility. 10thEuropean Congress of Endocrinology. Berlin, Germany, EndocrineAbstracts. 16: P613; Karaca, Z. and F. Kelestimur (2011), “Pregnancy andother pituitary disorders (including hGH deficiency).” Best Pract ResClin Endocrinol Metab 25(6): 897-910; Magon, N., S. Singh, et al.(2011), “Growth hormone in male infertility.” Indian J Endocrinol Metab15 Suppl 3: S248-249; and Sakai, S., T. Wakasugi, et al. (2011),“Successful pregnancy and delivery in a patient with adult hGHdeficiency: role of hGH replacement therapy.” Endocr J 58(1): 65-68, thecontents of each of which are incorporated herein by reference.

Until recently, hGH was available only in expensive injectable forms,and benefits from the restoration of hGH levels available only to thosewith the ability to pay. Most recently substances that can trigger therelease of human growth hormone from an individual's own anteriorpituitary gland have become available. These are generically referred toas secretagogues. Secretagogues have the ability to restore hGH levels,potentially to the levels found in youth. See, e.g., “Grow Young WithhGH” by Dr. Ronald Klatz, President of the American Academy ofAnti-Aging, published in 1997 by Harper Collins.

HGH-deficient adults have marked reductions in lean body mass, andwithin months of hGH treatment, gains in lean body mass, skin thickness,and muscle mass are observed. (Cuneo R C et al. J Appl Physiol 1991;70:695-700; Cuneo R C et al. J Appl Physiol 1991; 70:688-694; Rudman Det al. N Engl J Med 1969; 280:1434-1438)

It is well-established that intravenous (IV) administration of someamino acids results in significant hGH secretion. Intravenous infusionof 183 mg of arginine/kg body weight in females increased hGHlevels >20-fold, and 30 g of arginine elevated serum hGH levels 8.6 foldin males. (Merimee T J et al. N Engl J Med 1969; 280:1434-1438;Alba-Roth J et al. J Clin Endocrinol Metab 1988; 67:1186-1189) Otheramino acids, such as methionine, phenylalanine, lysine, histidine, andornithine, have also led to marked increases in hGH. (Alba-Roth, Muller,Schopohl, & von Werder, 1988; Chromiak & Antonio, 2002; Gourmelen, M.,M. Donnadieu, et al. (1972) Ann Endocrinol (Paris) 33(5): 526-528)

Given the difficulties in IV administration of amino acids forwidespread use, interest in elucidating the hGH response to oral aminoacid supplements prompted testing of such supplements containing mainlyarginine, lysine, and ornithine at varying amounts. Yet the pronouncedvariability in results among these studies makes clear the complexitiesinvolved in the design of an effective supplement for supporting hGHlevels in the general public. (Suminski R R et al. Int J Sport Nutr1997; 7:48-60; Lambert M I et al. Int J Sport Nutr 1993; 3:298-305;Corpas E et al. J Gerontol 1993; 48:M128-M133; Isidori A et al. Curr MedRes Opin 1981; 7:475-481; Fogelholm G M et al. Int J Sport Nutr 1993;3:290-297; Chromiak J A, Antonio J. Nutrition 2002 July; 18(7-8):657-61)

Serum hGH levels differ in relation to various factors including age,gender, hormone status, and BMI. (Iranmanesh, Lizarralde, & Veldhuis,1991); (Chowen, Frago, & Argente, 2004) Growth hormone has been shown tobe important in multiple stages of pregnancy including early antralfollicle recruitment, subsequent follicular growth, and oocytematuration. Moreover, administration of growth hormone during theovarian stimulation phase of in vitro fertilization (IVF) cycles hasbeen shown to increase the probability of clinical pregnancy.

BRIEF SUMMARY OF THE DISCLOSURE

Described herein are nutritional supplements and methods for using thesame. In embodiments, the nutritional supplement may be an amino acidsecretagogue composition, which, when administered orally, stimulatesthe pituitary gland of a subject to release growth hormone (e.g., hGH),and results in the promotion or maintenance of fertility.

Some embodiments include an oral nutritional supplement that comprises,for example and without limitation: L-arginine, oxo-proline, L-lysine,and cysteine. Particular examples include an oral nutritional supplementthat consists essentially of L-arginine hydrochloride, oxo-proline,L-lysine hydrochloride, N-acetyl-L-cysteine, L-glutamine, andschizonepeta powder.

Certain embodiments herein include an oral nutritional supplement dosageform that consists of 0.86 mmol L-arginine; 1.32 mmol oxo-proline; 2.05mmol L-lysine; 1.53 μmol N-acetyl L-cysteine; 1.71 μmol L-glutamine; and125 μg schizonepeta (aerial parts) powder. This oral nutritionalsupplement is referred to herein as “SeroVital®.” SeroVital® may beorally administered in an amount of, for example, 2.9 grams (i.e., 4unit dosage forms) to a human being, so as to stimulate the release ofhGH and/or increase testosterone levels in the human being. Thenutritional supplement may be administered on a regular basis, such as aweekly or monthly intake at a dosage tailored to the subject's needs;i.e., the nutritional supplement may be administered regularly asmultiples (1×, 2×, etc.) of the structural units (pills, tablets,capsules, etc.) in accordance with the needs of the subject.

Some embodiments include a method for increasing growth hormone (e.g.,hGH) and therefore increasing fertility in a subject (e.g., a humansubject) that comprises administering (e.g., orally) a nutritionalsupplement to a subject to improve one or more objective health metrics,including for example and without limitation: increasing or enhancingpregnancy and live birth rate; increasing lean body mass; reducingobesity, adipose tissue mass, and anxiety; supporting weight loss;decreasing appetite and atrophic processes in skeletal muscle, liver,kidney, spleen, skin, and bone; and improving at least one of energy,endurance, sleep, metabolism, heart rate, blood pressure, cardiovascularhealth, sympathetic nervous activity, thyroid response, glucoseutilization, mental/cognitive function, reaction time, bone density,hair health and appearance, nail health and appearance, skin health andappearance, libido, and at least one androgen-mediated condition(s).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 includes a plot of growth hormone levels measured in subjectsafter administration of an exemplary supplement compared to a placebo;

FIG. 2 includes a scatter plot and linear regression analysis of thetime to fall asleep in subjects with continued use of an exemplarysupplement over time; and

FIG. 3 includes a scatter plot and linear regression analysis of thetime awake during sleep over time in subjects with continued use of anexemplary supplement.

DETAILED DESCRIPTION

The determination of an effective and safe oral functional blend thatstimulates hGH secretion in the general population is important, sinceathletes, entertainers, and now the general public seek effective hGHsupport supplements and understand hGH to have rejuvenating properties.Indeed, once partial to athletes and entertainers, the desire foreffective supplements to provoke hGH increases now extends to thegeneral public. Not only do they have a goal of building lean tissue andreducing fat, but also of improving/increasing fertility and providingother rejuvenating qualities that hGH is understood to provide. However,the literature on oral amino acids for use in stimulating hGH does notcontain clear evidence for an optimized oral amino acid-containing blendable to stimulate hGH in the general public, including both men andwomen of a wide age range.

Therapeutic use of hGH has been used for improving pregnancy outcomes.Indeed, growth hormone has been shown to be important in multiple stagesof pregnancy including early antral follicle recruitment, subsequentfollicular growth, and oocyte maturation, and administration of growthhormone during the ovarian stimulation phase of in vitro fertilization(IVF) cycles has been shown to increase the probability of clinicalpregnancy.

Embodiments herein provide a nutritional supplement for elevating growthhormone (e.g., hGH) and subsequently improving fertility, pregnancy, andlive birth rate outcomes. Particular embodiments provide an aminoacid-containing composition that is well tolerated, and may have theresult of increasing or elevating hGH release in those individuals whosehGH release rates have slowed as a function of increasing age, or thathave normal hGH levels but desire higher hGH levels. The compositions ofthe present disclosure also result in increasing or enhancing fertility.There is a need for a nutritional supplement that efficiently anddemonstrably enhances the production and effects of natural human growthhormone and fertility outcomes in individuals of the general population.

Some embodiments herein provide a nutritional supplement for use by ahuman being. In particular embodiments, the nutritional supplement is anamino acid secretagogue composition, which, when administered orally,may stimulate the pituitary gland to produce hGH, and may jointlypromote or maintain fertility.

Increased production of hGH may result in enhancement of fertility;inhibition of insulin depression; inhibition of hyperglycemia andincrease in insulin effectiveness; enhancement of fat conversion;lowering of cholesterol; and/or normalization of lipid balance. Inexamples, a supplement herein may function as a dietary supplement byassisting the body's own ability to secrete hGH naturally, and in amanner which is safe and effective. Such a supplement may provide growthhormone therapy in a more affordable manner than existing compositionsand methods, for example, injectable compositions.

Particular embodiments herein include an oral nutritional supplementthat comprises L-lysine, L-arginine, oxo-proline, and one of cysteineand glutamine. In some examples, a supplement herein may comprise bothcysteine and glutamine and/or schizonepeta powder. In particularexamples, a functional dosage includes the L-arginine at a level betweenabout 0.1-6 mmol and the oxo-proline between about 0.1-8 mmol, and/orthe L-lysine in an amount between about 0.1-12 mmol. In particularexamples, the cysteine and/or glutamine may be contained at a levelbetween about 0.001-6 mmol.

Cysteine may be present in a supplement according to particularembodiments as n-acetyl L-cysteine, and the glutamine may beL-glutamine. Amino acids in a nutritional supplement herein may bedelivered as non-toxic salts thereof, effective complexes thereof,stable chelates thereof, active esters thereof, functional derivativesthereof, and mixtures thereof which are effective to increase hGH levelsin a subject from the general population.

Particular embodiments herein include an oral nutritional supplementthat consists essentially of L-lysine (e.g., L-lysine HCl), L-arginine(e.g., L-arginine HCl), oxo-proline, N-acetyl-L-cysteine, L-glutamine,and schizonepeta (aerial parts) powder. In particular examples, afunctional dosage includes L-arginine at a level between about 0.1-6mmol and oxo-proline between about 0.1-8 mmol, and/or L-lysine in anamount between about 0.1-12 mmol. n-acetyl L-cysteine and/or L-glutaminemay be comprised in some exemplary supplements at a level between about0.001-6 mmol. In particular examples, a functional dosage includesL-arginine at a level between about 2.5-4.5 mmol and oxo-proline betweenabout 4-6 mmol, and/or L-lysine in an amount between about 7-9 mmol.N-acetyl L-cysteine and/or L-glutamine may be comprised in someexemplary supplements at a level between about 0.001-0.5 mmol.

Certain embodiments herein include an oral nutritional supplement dosageform that consists of 0.86 mmol L-arginine; 1.32 mmol oxo-proline; 2.05mmol L-lysine; 1.53 μmol N-acetyl L-cysteine; 1.71 μmol L-glutamine; and125 μg schizonepeta (aerial parts) powder (SeroVital®). Thus, an oralnutritional supplement dosage form may consist of 181.38 mg L-arginineHCl; 170.93 mg L-pyroglutamic acid; 374.83 mg L-lysine HCl; 0.25 mgN-acetyl L-cysteine USP; 0.25 mg L-glutamine; and 0.125 mg schizonepeta(aerial parts) powder, for example, in a capsule. In some examples, theoral nutritional supplement may be administered to a human being byorally administering four such dosage forms (i.e., 725.50 mg L-arginineHCl; 683.70 mg L-pyroglutamic acid; 1499.30 mg L-lysine HCl; 1.00 mgN-acetyl L-cysteine USP; 1.00 mg L-glutamine; and 0.50 mg schizonepeta(aerial parts) powder).

Some embodiments herein provide a method for increasing or enhancingfertility in humans, and increasing human growth hormone in humans thatcomprises orally administering a nutritional supplement for elevatinggrowth hormone release to a healthy human being. As used herein, a“healthy human being” refers to a human being having any age-relateddecline in hGH, excluding any physiological deficiency that is not agerelated. Particular embodiments include oral administration of anutritional supplement for elevating growth hormone release to a humanthat is at least 30 years old.

Other embodiments herein provide a method for increasing or enhancingfertility in humans, and increasing human growth hormone in humans thatcomprises orally administering a nutritional supplement for elevatinggrowth hormone release to a human being that has a deficiency of hGH, orany condition resulting in low fertility or infertility, such as humanshaving pituitary disorders, idiopathic normogonadotropic infertility,and/or low hGH.

A nutritional supplement for elevating growth hormone release may beorally administered to a human being to improve health, including by:increasing lean body mass; reducing obesity, adipose tissue mass, andanxiety; supporting weight loss; decreasing appetite and atrophicprocesses in skeletal muscle, liver, kidney, spleen, skin, and bone; andimproving energy, endurance, sleep, metabolism, heart rate, bloodpressure, cardiovascular health, sympathetic nervous activity, thyroidresponse, glucose utilization, mental/cognitive function, reaction time,bone density, hair health and appearance, nail health and appearance,skin health and appearance, and libido.

In certain examples, oral administration of a nutritional supplement forelevating growth hormone release may improve fertility. Oraladministration of a nutritional supplement for elevating growth hormonerelease may also improve wound healing, provide anti-aging skinproperties, reduce wrinkles, dark spots, discolorations, dullness,sagging, laxity, and thinning, and may further improve texture,luminosity, “lift,” tone, radiance, smoothness, uniformity, and youthfullook of skin.

A nutritional supplement for elevating growth hormone release may beorally administered to a human being to improve health by affecting oneor more condition(s) and/or disease(s) that depend upon androgenactivity. Such conditions and diseases include, for example and withoutlimitation: low fertility; infertility; maintenance of fertility;maintenance of muscle strength and function; reversal or prevention offrailty or age-related functional decline (ARFD) in the elderly (e.g.,sarcopenia); remediation of age-related decreased testosterone levels inmen; treatment of male menopause; treatment of hypogonadism; malehormone replacement; treatment of male and female sexual dysfunction(e.g., erectile dysfunction, decreased sex drive, sexual well-being,decreased libido); male and female contraception; hair loss; Reaven'sSyndrome; enhancement of bone and muscle performance/strength; treatmentof catabolic side effects of glucocorticoids; prevention and/ortreatment of reduced bone mass, density, and/or growth (e.g.,osteoporosis and osteopenia); treatment of chronic fatigue syndrome(CFS); chronic myalgia; treatment of acute fatigue syndrome and muscleloss following elective surgery; acceleration of wound healing;acceleration of bone fracture repair; prevention of post-surgicaladhesion formation; acceleration of tooth repair or growth; maintenanceof sensory function (e.g., hearing, sight, olfaction, and taste);treatment of periodontal disease; treatment of wasting secondary tofractures and wasting in connection with, for example, chronicobstructive pulmonary disease (COPD), chronic liver disease, AIDS,weightlessness, cancer cachexia, burn and trauma recovery, chroniccatabolic state, eating disorders, and chemotherapy; treatment ofcardiomyopathy; treatment of thrombocytopenia; treatment of one or moresymptoms of Crohn's disease; treatment of short bowel syndrome;treatment of irritable bowel syndrome; treatment of inflammatory boweldisease; treatment of ulcerative colitis; treatment of complicationsassociated with transplantation; treatment of physiological shortstature; treatment of obesity and growth retardation associated withobesity; treatment of anorexia (e.g., associated with cachexia or ging);treatment of hypercortisolism and Cushing's syndrome; treatment ofPaget's disease; treatment of osteoarthritis; induction of pulsatilegrowth hormone release; treatment of osteochondrodysplasias; treatmentof depression, nervousness, irritability, and stress; treatment ofreduced mental energy and low self-esteem (e.g.,motivation/assertiveness); improvement of cognitive function (e.g.,treatment of dementia, including Alzheimer's disease and short-termmemory loss); treatment of catabolism in connection with pulmonarydysfunction and ventilator dependency; treatment of cardiac dysfunction(e.g., associated with valvular disease, myocardial infarction, cardiachypertrophy or congestive heart failure); lowering of blood pressure;protection against ventricular dysfunction or prevention of reperfusionevents; treatment of adults in chronic dialysis; reversal or slowing ofthe catabolic state of aging; attenuation or reversal of proteincatabolic responses following trauma (e.g., reversal of the catabolicstate associated with surgery, congestive heart failure, cardiacmyopathy, burns, cancer, COPD, etc.); reduction of cachexia and proteinloss due to chronic illness such as cancer or AIDS; treatment ofhyperinsulinemia, including nesidioblastosis; treatment ofimmunosuppressed patients; treatment of wasting in connection withmultiple sclerosis or other neuro-degenerative disorders; promotion ofmyelin repair; maintenance of skin thickness; treatment of metabolichomeostasis and renal homeostasis (e.g., in the frail elderly);stimulation of osteoblasts, bone remodeling, and cartilage growth;regulation of food intake; treatment of insulin resistance, includingNIDDM; treatment of insulin resistance in the heart; improvement ofsleep quality and correction of the relative hyposomatotropism ofsenescence due to high increase in REM sleep and a decrease in REMlatency; treatment of hypothermia; treatment of congestive heartfailure; treatment of lipodystrophy; treatment of muscular atrophy;treatment of musculoskeletal impairment; improvement of the overallpulmonary function; treatment of sleep disorders; treatment of thecatabolic state of prolonged critical illness; treatment of hirsutism,acne, seborrhea, androgenic alopecia, anemia, hyperpilosity, benignprostate hypertrophy, adenomas, and neoplasias of the prostate (e.g.,advanced metastatic prostate cancer) and malignant tumor cells includingthe androgen receptor, such as is the case for breast, brain, skin,ovarian, bladder, lymphatic, liver, and kidney cancers; treatment ofcancers of the skin, pancreas, endometrium, lung and colon;osteosarcoma; hypercalcemia of malignancy; metastatic bone disease;treatment of spermatogenesis, endometriosis, and polycystic ovarysyndrome; counteracting preeclampsia, eclampsia of pregnancy and pretermlabor; treatment of premenstrual syndrome; and treatment of vaginaldryness.

A nutritional supplement for increasing fertility and/or elevatinggrowth hormone release may be used in combination with other hormoneand/or steroid modulating supplements to enhance the effect of thenutritional supplements disclosed herein.

In accordance with the “consist essentially of” and “consistingessentially of” language herein, a nutritional supplement for elevatinggrowth hormone release in some embodiments is essentially limited to theaforementioned ingredients, and does not include any additional activeingredients intended to add nutritional content (e.g., vitamins,minerals, etc.), but may include additional ingredients not intended toadd nutritional content, for example, ingredients intended to fulfill anon-nutritional purpose (e.g., coloring, fillers, flavoring, aningredient for maintaining the structural form, etc.).

Each ingredient of a nutritional supplement for increasing fertilityand/orelevating growth hormone release may be prepared in accordancewith any method known to one of ordinary skill in the art.Alternatively, each ingredient may be obtained in a fully prepared formfrom a commercially available source.

A nutritional supplement for increasing fertility and/or elevatinggrowth hormone release may be in any suitable oral administration form,including but not limited to: a chewable form, a liquid form, a sprayform, a capsule form, a suppository form, dissolvable wafer, and apowder form. In some embodiments, a dosage form of the nutritionalsupplement may be present in an amount of about 2.9 grams.

Irrespective of the structural form of the nutritional supplement forincreasing fertility and/or elevating growth hormone release, theingredients of the nutritional supplement may be distributedhomogeneously or non-homogeneously within the nutritional supplement.

A nutritional supplement for increasing fertility and/or elevatinggrowth hormone release may be ingested on a regular basis, such as adaily or weekly intake at a dosage tailored to an individual's needs,i.e., the nutritional supplement is to be taken regularly as multiples(1×, 2×, etc.) of the dosage form (e.g., pills, tablets, capsules, etc.)in accordance with the needs of the individual. For example, a seniorcitizen leading a sedentary life is likely to need higher daily dosesthan does a young person engaged in regular strenuous exercise (e.g., aweight lifter).

Alternatively, the nutritional supplement for increasing fertilityand/or elevating growth hormone release may be ingested on an as-neededbasis at a dosage tailored to the individual's needs. Medical ornutritional counseling may be beneficial for arriving at a desirable oroptimal dosage tailored to the individual's needs. The nutritionalsupplement may be administered, for example, from one to three timesdaily, or, by way of further example, the supplement may be administeredevery other day or once a week. In particular embodiments, thenutritional supplement may be administered on an empty stomach.

In certain embodiments, a nutritional supplement for increasingfertility and/or elevating growth hormone release comprises a particularcombination of types of amino acids, mass ranges, and specificformulations that have been selected to be synergistically balanced andof adequate quantity to achieve a desired physiological effect, e.g.,growth hormone release and increased fertility. Improper combinations ofthe same amino acids may be ineffective. The component amino acids maybe synergistic in the sense that several of them, when combinedtogether, synergistically stimulate the release of human growth hormone,leading to enhanced fertility. The combination of amino acids inparticular embodiments was also chosen to reduce or inhibit chemicalcombination or reaction between the component amino acids.

EXAMPLES Example 1 Effect of an Oral Nutritional Supplement Single Doseon hGH Levels

The short-term effects of a single oral nutritional supplement on hGHlevels 2 hours post ingestion was studied in 16 healthy subjects [12males, four females; nine Caucasian, six African American, one other;mean age=32±14 years; body mass index=26.4±5.0 ranging from 19.1 to 36.8kg/m²]. Each subject reported to the Inpatient Unit on two occasions oneweek apart. After an overnight fast, subjects had an IV line placed andbaseline bloods samples were drawn at −30, −15,and 0 minutes.

Subjects were then asked to swallow the capsules of supplement(SeroVital®) or an identical looking placebo. SeroVital® is a 2.9 g/doseblend of L-lysine HCl, L-arginine HCL, oxo-proline, N-acetyl-L-cysteine,L-glutamine, and schizonepeta (aerial parts) powder. Blood was drawn at15, 30, 60, 90, and 120 minutes for assay. Human GH was measured at eachtime point using the Siemens Immulite™ 2000 (intra-assay CV was 3.72%,inter-assay CV was 5.70%, and the detection limit for hGH was 0.05ng/mL. The −15 and 120 minute time points were additionally assayed fortriiodothyronine (T3) as informative for mechanistic investigations.

The mean growth hormone increased 682% after the supplement from 0.17 atbaseline to 1.33 ng/mL at 120 minutes, compared to a mean decrease of52% after the placebo from 0.93 to 0.45 ng/mL. FIG. 1.

The mean change in hGH levels from baseline to 120 minutes (hGH at 120minutes minus hGH at 0 minutes), was 1.15 (95% CI: 0.17, 2.14) ng/mLafter the supplement, versus −0.48 (−1.47, 0.50) ng/mL after theplacebo, demonstrating a statistically significant differential effect(P=0.01). After the supplement, the mean AUC for hGH across 120 minuteswas 20.43 (95% CI: 19.90, 20.95) ng/mL/min which was significantlyhigher (P=0.04) than placebo at 19.67 (18.74, 20.59) ng/mL/min. Overall,120 minutes after taking the supplement, hGH levels were significantlyhigher in absolute levels or by AUC.

As daily circadian levels of T3 naturally decrease during the morninghours, at which the current trial was scheduled, it was not surprisingthat placebo levels between the −15 and 120 minute time points decreasedby −6.10 ng/dL (106 to 100 ng/dL, P=0.01). In contrast, the SeroVital®group exhibited a deceased reduction in T3 by nearly one-half over thesame time course, −3.3 ng/dL (101-97.3 ng/dL, NS), which was not asignificant reduction compared to baseline, as was the reduction in theplacebo group. These results affirm that somatostatin inhibition plays amechanistic role in the ability of SeroVital® to induce significantincreases in serum hGH levels in human subjects.

At 120 minutes, hGH concentrations were two-fold higher in women(2.3±1.1 ng/mL, n=4) than in men (1.0±0.4 ng/mL, n=12, although thestudy was not adequately powered for these comparisons. Nevertheless,these findings support an enhanced effect of the SeroVital® supplementin women.

An eight-fold increase was observed, equivalent to 682%, in hGH levels120 minutes after a single oral supplement of SeroVital®. The study hada broad range of ages and BMIs and included both genders. An additionaladvantage of the present study over previous hGH evaluations is that itcontained a placebo control group and was randomized and double-blinded.

These findings demonstrate that a specialized low-dose amino acidsupplement can significantly increase short-term hGH levels. Futurestudies will examine whether such increases in hGH with oral amino acidsupplementation increase fat-free mass and strength. This indeed may bethe case, since elderly subjects administered oral hGH secretagogues forsix and 12 months have sustained increases in lean body mass andimproved physical function.

The absolute magnitudes of these results are somewhat difficult todirectly correlate among past studies, as commercial hGH assays usedifferent antibodies to target specific hGH epitopes. Therefore,different antibodies and assays are less likely to recognize somespecific isoforms and fragments of the hGH molecule. This results invariability of the normal range of the hGH measurements in differentassays. Indeed, the same hGH sample measured using different assays canvary two- to three-fold, limiting the ability to compare actual hGHlevels across studies. Nevertheless, the mean levels of hGH reachedafter the subcutaneous injection of 0.06 IU of hGH in the treatment ofhGH deficient subjects was 0.4 ng/mL, a value that was clearly in therange of values seen in our study with oral amino acids.

Findings obtained from a randomized, blinded, placebo-controlled studystrengthen the evidence that oral administration of amino acids, whencompounded properly, can increase hGH serum levels, wherein SeroVital®administration showed an eight-fold increase, equivalent to a 682%increase in hGH levels, 120 minutes after a single oral dose. Inaddition, we elucidate some mechanistic details for these significanthGH increases as through somatostatin inhibition, supported by ourresults on the 120 minute results on T3 levels in the same subjects.

Example 2 Effect of an Oral Nutritional Supplement on Fertility

The effects of an oral nutritional supplement on fertility were studiedin a group of healthy human subjects. Subjects were asked to swallowcapsules of SeroVital®.

Case 1: A 27-year-old woman was admitted to St. Mark's Center forWomen's Health for a physical examination with complaints of irregularmenses and secondary infertility. Six years previously, she had givenbirth to her first child. The pregnancy was uneventful with spontaneouslabor at 38 weeks. She underwent cesarean section for delivery afterfailure to progress in labor, and gave birth to a healthy baby. Thesubject was unable to breastfeed because she did not have sufficientmilk production.

Following the birth of her first child, she had experienced weight gainand difficulty losing weight. Her menstrual periods were light andinfrequent, numbering approximately two per year with each lastingapproximately three days. She and her partner failed to conceive apregnancy over a six-year period while actively attempting to getpregnant for the last four years.

Upon examination, clinical results were consistent with Polycystic OvarySyndrome (PCOS), and a uterine ultrasound was performed. Ultrasoundconfirmed polycystic ovaries and PCOS diagnosis. The patient elected aconservative mode of management by engaging in a targeted diet andexercise program for weight loss. However, she was only able to lose amarginal amount of weight and experienced no changes in menstrualcycles.

After approximately a year and a half of attempting weight loss as theonly mode of management, the subject started daily supplementation withSeroVital® (2.9 g/day). After the first two months of SeroVital® intake,she had a menstrual period. After one more month of supplementation, shehad another menstrual period. After one more month of supplementationfor a total of four months of supplementation, she confirmed pregnancy.She discontinued SeroVital® supplementation after pregnancy wasconfirmed.

Her second pregnancy proceeded to term without complications and thepatient gave birth to a healthy baby boy weighing 10 lbs 6 oz, length21½ inches.

Case 2: A 33-year-old woman, BMI 37.2, was referred to a reproductivecenter on Aug. 15 2013 for secondary infertility and recurrent pregnancyloss. Medical impression included SAB×2 (1 with prior partner); SVD×1;ovulatory dysfunction; menses every 2-3 months; moderate dysmenorrhea;minimal endometriosis. [Male factors showed normal semen analysis].

History: In May of 2001 she experienced an SAB (miscarriage). In Januaryof 2009, she gave birth to a healthy daughter (preeclampsia, on bedrestfrom 29 weeks), having conceived after a second cycle of Clomid. InJanuary of 2012, she experienced a second SAB (bleeding×2 weeks, serumHCG low and then went down, no sac seen on ultrasound, biochemical).

In August of 2013, the patient had her initial consult and initiatedtreatment:

Ovulation Induction (04 anovulation

The patient presented with the following test results and treatment:

-   -   28 Aug. 2013 Baseline US showed 19 total antral follicle counts        (AFC), 4 mm endometrial thickness, 1.5 cm leiomyoma in anterior        wall of fundus noted    -   28 Aug. 2013 E2<25, antimullerian hormone (AMR) 0.8, follicle        stimulating hormone (FSH) 2.4, Prolactin 16.4, HgbAlc 5.5%, TSH        1.2    -   Femara 5 mg days 3-7    -   13 Sep. 2013 P4 (progesterone) 0.2, day 23    -   Provera 10 mg×10 days

Intrauterine Insemination (UI), Conceived

The patient presented with the following test results and treatment:

-   -   21 Nov. 2013 Menses    -   Femara 7.5 mg days 3-7    -   Took SeroVital® daily from 29 Nov. 2013 to 6 Jan. 2014    -   5 Dec. 2013 ultrasound on day 14 showed 15 total AFC, lead        follicle measuring 23 mm on the right, 7.5 mm endometrial        thickness    -   Human chorionic gonadotropin (HCG) 10k    -   7 Dec. 2013 IUI on cycle day 15. Semen sample showed 4.7 ml, 109        mill/ml, 67% motility, 343.2 mill total motile sperm count        (TMC), 2.5 progression preprep    -   21.9 mill TMC inseminated after gradient preparation    -   30 Dec. 2013 HCG 5921    -   2 Jan. 2014 ultrasound showed single IUP measuring 5 weeks 6        days, FHT 114

Case 3: A 29-year-old woman, 156 cm tall, weighing 49.9 kg (BMI 20.5kg/m2) desired a pregnancy but had been impacted by endometriosissymptomology since adolescence. She declined surgical and hormonalinterventions since her initial diagnosis and managed symptoms ofmenorrhagia and dysmenorrhea through nutritional and active-lifestyleapproaches.

After six months of unprotected intercourse, the subject conceived. Shegained 29 kg over the course of pregnancy and suffered from mild nausea,occasional vomiting, and persistent colds. She otherwise had anon-eventful pregnancy and underwent a planned cesarean section forbreach presentation at 39 weeks. After delivery, the subject was able tobreastfeed for several weeks, after which she did not have sufficientmilk production to continue breastfeeding. The subject experienced sharpabdominal pains with any physical motion at the site of cesarean sectionincision for several months after the delivery and was unable toexercise until the pain subsided. The subject also experienced symptomsof postpartum depression.

Four months following the delivery, the subject began dailysupplementation with SeroVital® (2.9 mg/day) and engaged in fitnesstraining. She lost 25 kg and competed in a competitive running event.Nine months after beginning daily supplementation with SeroVital®, thesubject conceived an unplanned, second pregnancy from a singleunprotected event. She discontinued SeroVital® supplementation afterpregnancy was confirmed.

The subject suffered from increased symptoms of illness compared to herfirst pregnancy including nausea, vomiting on 12 occasions, andpersistent colds, but had no other complications. She underwent aplanned cesarean section at 42 weeks after spontaneous labor was notreached. The subject breastfed her baby for a period of two weeks. Sheinitiated daily SeroVital® supplementation four weeks after the deliveryand two weeks after ceasing breastfeeding.

Three months after restarting daily supplementation with SeroVital®, thesubject completed a half-marathon. One month later and four months afterrestarting SeroVital®, the subject conceived an unplanned, thirdpregnancy, once again from a single unprotected event. The subjectcontinued SeroVital® supplementation throughout the pregnancy and didnot experience symptoms of illness as she did with her previouspregnancies. She underwent a planned cesarean section and tubal ligationat 39 weeks, giving birth to her third healthy baby.

As no other observable changes in diet or medication took place,SeroVital® appears to have heightened her fertility, leading to hertimely second and third improbable pregnancies. Ingestion of SeroVital®also appears to have reduced postpartum depressive symptoms after hersecond and third deliveries, as well as eliminating physical illnesssymptoms during her third pregnancy. She notes that she did notexperience pain with physical motion at the incision site after hersecond and third cesarean sections as she did with her first, despitethe short succession between multiple surgeries.

Example 3 Effect of an Oral Nutritional Supplement on Endurance and FatMetabolism

To test the effect of the nutritional supplement on endurance and fatmetabolism, a double-blind clinical study was conducted involving 12healthy subjects [seven males, five females; mean age=31±6 years; bodymass index=25.7±3.8 ranging from 20.3 to 32.2 kg/m²]. Each subjectreported to the Fitness Testing Facility (PEAK, University of UtahCollege of Health) after an overnight fast. Upon arrival, each subjectunderwent standard measurements of weight, height, body fat percent (BodPod), and resting metabolic rate (RMR, indirect calorimetry). Dailycalorie expenditure was estimated based on the additive evaluations ofmeasured RMR, estimated Lifestyle and Activity (defined as the number ofcalories burned performing daily activities including working, playing,eating, etc.), and estimated Exercise (defined as an estimate of thenumber of calories burned during exercise based on daily activitylevel). Following the baseline measurements, subjects then consumed astandard breakfast (Egg McMuffin, 300 calories; 12 g fat; 29 gcarbohydrates; 18 g protein). Subjects rested for a further 45 minutesto reach a post-absorptive state, then underwent a Maximal AerobicFitness Test of graded exercise, completed on a treadmill. Subjects'oxygen uptake was measured using a metabolic cart, and VO_(2max) wasquantified.

Subjects were then provided a two-week supply of a novel supplementSeroVital® (2.9 g/dose blend of L-lysine HCl, L-arginine HCl,oxo-proline, N-acetyl-L-cysteine, L-glutamine, and schizonepeta (aerialparts) powder). The novel SeroVital® increases serum human growthhormone hGH levels by eight times (equivalent to 682%) 120 minutes aftera single dose in healthy male and female volunteers. In this study,subjects were instructed to consume one dose of the supplement on anempty stomach, two hours after dinner prior to bedtime, every night forthe two-week study duration.

Following the final dose, each subject returned to the PEAK FitnessTesting Facility, University of Utah College of Health, after anovernight fast (without having consumed their last dose of thesupplement since the previous night). Upon arrival, each subjectunderwent the identical test protocols as the baseline test day. Thedata from the two measurement days were then analyzed. Consistent withthe hypothesis that the supplement would improve endurance parameterswith its ability to increase in hGH levels, the decision was made toreject the null only if the data supported the one directionalalternative consistent with a favorable response to the supplement.Statistical significance was assumed for P≦0.05.

Mean VO_(2max) increased by 6% from 44.9±8.1 at baseline to 47.7±9.2mL/kg/min (3.69±0.96 to 3.91±1.02 L/min), demonstrating a statisticallysignificant differential effect compared to baseline (P=0.02). After theperiod of supplementation with SeroVital®, the mean RMR increased by2.7% from 1687±330 to 1733±288 kcal/day with a statistical trend towardssignificance compared to baseline (P=0.165). Estimated daily calorieexpenditure also increased by 2.7% from 1687±330 to 1733±288 kcal/daywith the trend towards significance (P=0.166).

After two weeks of supplementation with the supplement SeroVital® (takenon an empty stomach, two hours after dinner prior to bedtime, everynight), both RMR and estimated daily calorie expenditure tended toincrease, evidencing the potential of the supplement to impart long-termfat burning effects. Additionally, endurance as measured by VO_(2max) inthe post-absorptive state significantly improved with a measured 6%increase. Overall, the SeroVital® supplement was shown to increaseparameters of endurance, energy, and vitality.

Example 4 Effect of an Oral Nutritional Supplement on Sleep Improvement

To test the ability of the nutritional supplement to improve sleep, adouble-blind clinical study was conducted involving 15 healthy subjects[10 males, five females; mean age=33±7 years]. Each subject completed abaseline Epworth Sleepiness Scale self-report questionnaire and astandardized assay of usual sleep habits. All subjects were deemed tohave average sleep parameters within a normal range.

The subjects were then provided a three-week supply of a novelsupplement SeroVital® (2.9 g/dose blend of L-lysine HCl, L-arginine HCl,oxo-proline, N-acetyl-L-cysteine, L-glutamine, and schizonepeta (aerialparts) powder). The novel SeroVital® blend increases serum human growthhormone hGH levels by eight times (equivalent to 682%) 120 minutes aftera single dose in healthy male and female volunteers.

We investigated sleep patterns with continued use of the supplement whentaken on an empty stomach, two hours after dinner prior to bedtime,every night for three weeks. On each trial day, subjects reported 1)time went to bed; 2) time of final awakening; 3) estimated time to fallasleep; 4) time of awakening during sleep/length of time awake. Data wascompiled by day for estimated time to fall asleep and length of timeawake during sleep in order to assess sleep efficiency. Daily values foreach measure were plotted as an average (±S.D.) among the subjects overthe time course of the study, and a linear regression was tabulated toassess overall trends over time. All available data was included in theanalysis.

Linear regression analysis showed that both estimated time to fallasleep (FIG. 2) and time awake during sleep (FIG. 3) tended to decreaseover time with continued use of the supplement over the time course ofthe study. The time to fall asleep decreased with an average slope of−0.24 min/day, and the time awake during sleep decreased by an averageslope of −0.26 min/day. Overall, these results show a trend towardsgreater sleep efficiency by measurements of both time to fall asleep andtime awake during sleep, both with a quantified average decrease ofabout 0.25 min/day over three weeks with regular nighttime use of thenovel SeroVital® supplement (when taken as directed, on an emptystomach, two hours after dinner prior to bedtime).

Example 5 Effect of an Oral Nutritional Supplement on Lean Body Mass andWeight Change

To test the effect of the nutritional supplement on lean body mass andweight change of humans, healthy men and women between the ages of 30 to80 years are recruited through advertisements. Standardized assessmentcriteria are used to select subjects at risk for functional decline(e.g., hand grip strength, habitual gait speed, etc.). Additionally,subjects are excluded for diabetes mellitus, use of anticoagulants,seizure disorder, cancer treatment within five years, poorly controlledhypertension, unstable or recent onset angina, myocardial infarctionwithin six months, cognitive impairment, depression, significantlimitations of lower extremity function, bradycardia, systolic bloodpressure <100 or >170 mm Hg, or orthostatic hypotension. Subjectsparticipating in strength training programs were also excluded.

Subjects are given daily doses of the nutritional supplement for sixmonths at different dosing concentrations of active ingredients, and onegroup is treated as a placebo group. The study includes a pre-screeningassessment, screening and baseline visits, and weekly visits over theplanned six months, where weight, body composition, and physicalperformance (including endurance) are determined, along with hormonaldata analysis. Weight, percent lean body mass, and percent fat body massare chosen as primary measures of body composition at the six-monthanalysis period.

Example 6 Effect of an Oral Nutritional Supplement on Energy, BoneDensity, Skin Thickness, and Mass of Adipose Tissue

To test the effect of the nutritional supplement on energy, bonedensity, skin thickness, and mass of adipose tissue, healthy men over 60years old are recruited through advertisements. Standardized assessmentcriteria are used to select subjects at risk for functional decline(e.g., hand grip strength, habitual gait speed, etc.). Entry criteriainclude body weight of 90 to 120 percent of the standard for age.Additionally, subjects are excluded for diabetes mellitus, use ofanticoagulants, seizure disorder, cancer treatment within five years,poorly controlled hypertension, unstable or recent onset angina,myocardial infarction within six months, cognitive impairment,depression, significant limitations of lower extremity function,bradycardia, systolic blood pressure <100 or >170 mm Hg, or orthostatichypotension. Subjects participating in strength-training programs werealso excluded.

Subjects are given daily doses of the nutritional supplement for sixmonths at different dosing concentrations of active ingredients, and onegroup is treated as a placebo group. The study includes a pre-screeningassessment, screening and baseline visits, and weekly visits over theplanned six months, where lean body mass, mass of adipose tissue, skinthickness, and bone density at various skeletal sites are measured.Subjects are also monitored for changes in energy levels throughout theexperiment.

Example 7 Effect of an Oral Nutritional Supplement on Anxiety and Sleep

To test the effect of the nutritional supplement on anxiety and sleep,healthy men and women between the ages of 30 to 80 years are recruitedthrough advertisements. Subjects are interviewed by a psychiatrist usingthe Structured Clinical Interview for DSM-III-R or analogous method.Patients are all drug free. The study focuses on the evaluation ofanxiety, sleep patterns, and measurement of basal morning stress hormonelevels at various points during the six-month testing period.

Subjects are given daily doses of the nutritional supplement for sixmonths at different dosing concentrations of active ingredients, and onegroup are treated as a placebo group. The study includes a pre-screeningassessment, screening and baseline visits, and weekly visits over theplanned six months, where anxiety assessment (e.g., Hamilton AnxietyRating Scale) and anxiety and stress hormone levels (e.g., cortisol andhGH) are determined, and sleep study analysis are conducted.

Example 8 Effect of an Oral Nutritional Supplement on TestosteroneLevels

The effects of an oral nutritional supplement on testosterone levels arestudied in a group of healthy human subjects. Subjects are asked toswallow the capsules of SeroVital® or an identical looking placebo.

Blood samples (8-10 mL) are taken from each participant before use,after six days of use, after 12 days of use, and three days aftersuspending use. Each blood draw is taken in the morning between 9:30 and10:30 am, at a time when serum oscillations of testosterone is at itsminimum daily value. The determination of testosterone in human blood iscarried out using commercially available immunochemoluminescence assays.(Topo et al., 2009)

Results on testosterone release show that after using the SeroVital®supplement, serum testosterone levels are increased in a significantnumber of the subjects taking the supplement. On average, the increasein serum testosterone is statistically significant compared to those ofthe placebo group over the time course of the study.

While embodiments of the present invention have been described hereinfor purposes of illustration, many modifications and changes will becomeapparent to those skilled in the art. Accordingly, the appended claimsare intended to encompass all such modifications and changes as fallwithin the true spirit and scope of this invention.

We claim:
 1. A method for increasing, enhancing, or maintainingfertility in a human being in need thereof, the method comprising:providing an oral nutritional supplement unit dosage form, wherein theoral nutritional supplement unit dosage form comprises about 1 mmolL-arginine, about 1 mmol oxo-proline, about 2 mmol L-lysine, about 1.5μmol N-acetyl L-cysteine, about 2 μmol L-glutamine, and about 125 μgschizonepeta (aerial parts) powder orally administering an effectiveamount of the nutritional supplement to the human being.
 3. The methodaccording to claim 1, wherein the oral nutritional supplement unitdosage form comprises: about 0.9 mmol L-arginine; about 1.3 mmoloxo-proline; about 2.0 mmol L-lysine; about 1.5 μmol N-acetylL-cysteine; about 1.7 μmol L-glutamine; and about 1.25 μg schizonepeta(aerial parts) powder.
 4. The method according to claim 3, wherein theoral nutritional supplement unit dosage form comprises about 181 mgL-arginine HCl; about 170 mg L-pyroglutamic acid; about 374 mg L-lysineHCl; about 0.25 mg N-acetyl L-cysteine USP; about 0.25 mg L-glutamine,and about 0.125 mg schizonepeta (aerial parts) powder.
 6. The methodaccording to claim 1, wherein the oral nutritional supplement unitdosage form is administered to the human being to further: increasepregnancy rate; increase live birth rate; increase lean body mass orlibido; reduce obesity, adipose tissue mass, or anxiety; support weightloss; decrease appetite and atrophic processes in skeletal muscle,liver, kidney, spleen, skin, or bone; improve energy, endurance, sleep,metabolism, heart rate, blood pressure, cardiovascular health,sympathetic nervous activity, thyroid response, fat metabolism, glucoseutilization, mental/cognitive function, reaction time, bone density,hair health and appearance, nail health and appearance, skin health andappearance, or libido; and/or affect one or more condition(s) and/ordisease(s) that depend upon androgen activity in the human being.
 7. Themethod according to claim 1, wherein the oral nutritional supplementunit dosage form is orally administered in a total amount of about 2.9grams.
 8. The method according to claim 1, wherein the oral nutritionalsupplement is administered from one to three times daily.
 9. The methodaccording to claim 1, wherein the oral nutritional supplement isadministered once a week.
 10. The method according to claim 1, whereinthe human being is at least 30 years old.
 11. The method according toclaim 1, wherein the human being exhibits one or more symptoms of hGHdeficiency.